title: Detection of ABCC2, CYP2B6 and CYP3A4 in human peripheral blood mononuclear cells using flow cytometry.
abstract: ABCC2 has a wide tissue distribution and can mediate the efflux of a number of therapeutic compounds from cells and contribute to potential treatment failure. Its diverse expression and ability to efflux a number of substrates imply a number of physiological and pharmacological roles. CYP2B6 and CYP3A4 are responsible for the metabolism of a number of therapeutic compounds. Reports on the expression of these proteins in various cells and tissues have been contradictory mainly due to differences in experimental approach and cell type studied. With the advances in commercially available antibodies we describe here a simplified technique for the detection of ABCC2, CYP2B6 and CYP3A4 in human peripheral blood mononuclear cells (PBMC) by flow cytometry. Results are expressed as mean increase in fluorescence compared to isotypically matched controls. Using these assays we confirmed the expression of these proteins in human PBMC. These methods are rapid and reproducible and have potential use for both in vitro and clinical applications.
authors.affiliation: Department of Pharmacology and Therapeutics, The University of Liverpool, 70 Pembroke Place, Liverpool, L69 3GF, UK. neill.liptrott@liv.ac.uk
authors.forename: N J
authors.initials: NJ
authors.lastname: Liptrott
authors.forename: S H
authors.initials: SH
authors.lastname: Khoo
authors.forename: D J
authors.initials: DJ
authors.lastname: Back
authors.forename: A
authors.initials: A
authors.lastname: Owen
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis*; D065702:Cytochrome P-450 CYP2B6; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis*; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis*; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis*
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; D065702:Cytochrome P-450 CYP2B6; D051544:Cytochrome P-450 CYP3A; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D010089:Oxidoreductases, N-Demethylating; C000715087:ABCC2 protein, human; C585599:CYP2B6 protein, human; C510163:CYP3A4 protein, human